Podium Abstracts

Milestones and Scopes of Activity in General and Applied Microbiology in Armenia

The long-term activity of the lead institutions in general and applied microbiology in Armenia will be presented. Data obtained provide high efficiency of Spirulina microalgae for production of food and feed products on soda-salted soils of Ararat valley. The Culture Collection of entomopathogenic bacilli with over 5000 strains isolated from different continents all over the world has been established. On the industrial scale production of new bacterial insecticides has been organized. Perspectives for obtaining of cancerolytic preparations, novel Parasporins from crystalline inclusions producers, are discussed. As novel and prospective direction the microbiological production of biofuels on renewable sources and related topics have been emphasized.

About half of the ~200 virus resistance genes in plants are recessively inherited, suggesting that recessive resistance is more common for viruses than for other plant pathogens. The use of such genes is therefore a very important tool in breeding programs to control plant diseases caused by pathogenic viruses. This type of resistance is preferentially expressed in protoplasts and inoculated leaves, influencing virus multiplication at the single-cell level or cell-to-cell movement. Molecular mechanisms of recessive resistance will be described and discussed in this seminar based on our experience with cucurbits and cucurbit viruses. Cucurbit crops are affected by a significant number of virus-induced diseases which can drastically reduce yield and fruit quality. Sources of genetic resistance to cucurbit viruses have been identified in a number of cucurbit accessions, and some of them are being exploited commercially. These include the melon recessive nsv gene, a natural source of resistance to Melon necrotic spot virus (MNSV). The nsv locus codes for an allele of the eukaryotic translation initiation factor 4E (eIF4E). Genetic evidence suggests that an interaction occurs between eIF4E and a 3?-cap-independent translational enhancer located in a non-coding region of the virus genome. The specificity of this interaction appears to regulate MNSV host range, thus controlling MNSV susceptibility/resistance in melon. In agreement with the description of eIF4E as a factor required for virus multiplication in several virus/host combinations, eIF4E knocked-down melon plants show broad range virus resistance. While the recessive nsv resistance fits into the mechanistic model of passive resistance (i.e., lack of susceptibility), other recessive resistances in melon seem to act differently. These include resistance shown by accession TGR-1551 to Watermelon mosaic virus (WMV). In this case, high virus multiplication is allowed in initially inoculated tissues, but a restriction to virus accumulation in whole plants occurs afterwards. Microarray analyses supported the hypothesis that induction of a resistance response is restricting WMV colonization of resistant TGR-1551 plants.

The 36-month BIOPARTNERS project gives Georgia the possibility to improve the research activities of their highest quality in the field of food and biotechnology, the project includes several types of activities (Twining activities, exchange of researchers…).

Horizon 2020 will give new opportunities, and is fully open to Georgian participation in all fields and areas. Georgian researchers are able to team up with their European partners to participate in projects. In addition, a number of specific calls are targeting cooperation with Georgia, notably in the areas of Food, Agriculture and Biotechnology.

Pomegranate (Punica granatum L.) has become to be an attractive product for consumers both for its organoleptic properties as its high content of phenolic compounds. The aim of this conference is to summarize some technological development of products with commercial interest and high value-added products for the integral use of this fruit.

The use of different processing technologies to obtain industrial juice from Spanish pomegranate cultivars was evaluated. Furthermore, the extraction efficiency of pomegranate juice using industrial blender, press or turbo finisher in presence or not of peel was tested in order to obtain a juice rich in bioactive compounds. The effect of different stabilization technologies of the juice on its physicochemical composition, focusing on compounds with biological activity, as well as colour and turbidity, when subjected to heat treatments was also evaluated. The juice was treated from 65°C 30s to 90°C 5s (time / temperature conditions) to compare the effectiveness of different heat treatment concluding that low pasteurization temperature is able to reduce the initial microbial load of the juice up to safe values (more than 5 log reduction in cfu of the initial population) with a lower degradation of colour and bioactive compounds.

Finally current situation of functional drinks and pomegranate wine development as well as guidelines for nutraceutical production from peel and seeds will be analyzed.

Submerged cultivation of medicinal fungi biomass and its active pharmaceutical compounds, in large scale, is one of the most used technologies. The mechanism of fungal growth on solid state substrates is strongly connected pasting the fungal tips on the solid matrix. For this purpose fungal polysaccharides are producing and accumulating on the tip of young hyphae. In further process this jelly material serve as the sticky plaster for anchoranging the fungal tips on the solid matrix. Polysaccharide gel serve also as the matrix for the transfer of cellulolytic and xylolytic enzymes on the surface of cellulose material. On solid matrix fungal mycellia use polysaccharide gels also as a basis for moving on the solid matrix.

In submerged cultivation accumulation of fungal polysaccharides according to the primary needs is not needed. In high shear field is peeling polysaccharide gel cover from of the tips of young hyphae in the liquid substrate increasing the pseudoplasticity of submerged cultivation broth. Increased pseudoplasticity of the cultivation broth rapidly reduces mass and heat transfer. Oxygen transfer is limited and higher agitation and more intensive aeration is requested. Increased shear forces in liquid media could influence on further fungal growth.

The other problem is exposed with foaming and flotation of fungal biomass where the tips of young hyphen use polysaccharide cover gel for gluing the tips on bioreactor was anchoranging fungal biomass on the bioreactor head space. In further, fungal biomass could cover also all of the sensors and measuring devices that could change the quality of the signal very drastically.

Solid state cultivation of fungal biomass on solid matrix is one of the solutions of this problem. The benefits and the differences of both technologies as well as the economics would be discussed.

Recent Results on Foodomics Investigations of Cancer and Metabolomics of Alzheimer: New Advances to Study the Bioactivity of Dietary Ingredients

We will present some of the latest Foodomics results obtained in our laboratory. Namely, these Foodomics works were done: i) to investigate the possibilities of Foodomics in Alzheimer’s disease studies and, ii) to determine the anti-proliferative effect of food ingredients against different human cancer cell lines. Whole-transcriptome microarray, proteomics and MS-based non-targeted whole-metabolome approaches were employed to carry out the mentioned studies. These Foodomics strategies enabled: i) the identification of biomarkers for early detection of Alzheimer’s disease which should allow to investigate the effect of diet on this illness, and ii) the identification of several differentially expressed genes alone and/or linked to changed metabolic pathways that were modulated by food ingredients in cancer cells, providing new evidences at molecular level on the antiproliferative effect of food compounds.

Problems on Commercialization of Agricultural Biotechnology in Georgia

The agriculture is extremely important to the economy of Georgia and has a direct effect on the lives of the majority of Georgians, but after the restoration of independence the agricultural and food sector has undergone a deep crisis and the country of agricultural net-export changed into country of net-import. Development of agricultural biotechnology can contribute to the economic growth of the country by commercializing plant cultivars that possess improved genetic characteristics. Despite increasing use of the modern agricultural biotechnology elsewhere, efforts to put these inventions and launch profitable products in Georgia were unsuccessful due to protests from the public and politicians against genetically modified plants.

The importance of biosafety in Georgia is conditioned by a number of factors, particularly, due to the diversity of altitudes, soil-climatic zones and landscapes, the country hosts a high diversity of plants and animals, including many endemic species. Georgia is considered as important center of agrobiodiversity in the world, and represents a relatively large area supporting wild relatives of agricultural varieties. In this context, the authorities have developed a complex legislation for agricultural biotechnology that limits research, development, production and imports. The scientific discussions can lead to constructive ideas on how to resolve some of biotechnology’s most pressing issues and allow safe applications to be commercialized in Georgia. The report summarizes biotechnology commercialization in the world, marks significant agricultural biotechnologies and accents results in biotechnology commercialization in Georgia. The analysis has been made by using various sources of agriculture and biotechnology statistics, and the focus of the analysis is on analyzing the factors behind the concept “quality and competitiveness”.

Foodomics, defined for the first time in our research group attempts to provide a global vision on the relationship between food and health through the use of -omics technologies with the mail goal of improving consumer’s well-being, health and confidence. Green Foodomics attempts to contribute to the greenness, sustainability and ecology of Foodomics as a whole.

Foodomics can be seen as a green discipline trying to provide with new answers to the challenges of our society; aspects related to the sustainability, food quality and safety are basically embedded in the own Fodomics’ definition. Other aspects such as those related with the rational design of new foods able to improve human health and to prevent illnesses are basically green by themselves since they will contribute to obtaining safer foods, with lower contamination and chemical risks.

In this presentation, different green alternatives for the production of new functional food ingredients (based on the use of green solvents and the design of integrated processes producing less residues and consuming lower amounts of energy) will be presented together with the possibility of applying those processes to develop greener analytical methodologies to face some aspects related to the food quality, traceability and safety (through the miniaturization of sample preparation techniques, the use of ecological solvents and the development of new separation methods).

Recently, fundamental and applied research on bioconversion of lignocellulosic biomass by higher basidiomycetes has markedly increased. Because of the renewable nature and abundant availability, plant biomass has attracted considerable attention as alternative feedstock and potential chemicals and energy source. In this presentation, the state of the art of lignocellulose bioconversion into value-added products by submerged and solid-substrate fermentation will be presented. It integrates the recent data on the physiology of wood-rotting basidiomycetes, focusing on the common characteristics and unique properties of individual fungi as well as on several approaches providing efficient expression of their biosynthetic and destructive potential.

Olive oil press cakes (OOPC) represent a waste having a negative impact on environment. OOPC have little or no use and because of that solutions for their alternative use are sought. We investigated substrate mixtures composed of different proportions of OOPC, wheat bran, crushed corn seeds and beech sawdust for cultivation of Ganoderma lucidum, Lentinula edodes and Grifola frondosa fruiting bodies. Trend of fruiting bodies yields reduction was noticed, when substrates contained higher amounts of OOPC. In case of G. frondosa yields tend to reduce slower when instead of wheat bran crushed corn seeds were used as a substrate supplement. Results show that although OOPC in small portion can be successfully used as a medicinal mushroom fruiting bodies cultivating substrate, their use is rational only if no other substrate composing materials can be found.

CEBAS-CSIC is a research institute belonging to the Spanish National Research Council (CSIC), the largest public research organization in Spain and the third in Europe. CSIC’s mission is to generate knowledge through research, to train scientists and to apply its knowledge for the benefit of society. CSIC has 139 research institutes and centers nationwide and its activities are divided in 8 areas of knowledge. CEBAS-CSIC is one of the leading CSIC research centers in Agri-food Science & Technology. Since 2002, CEBAS-CSIC has an active technology transfer unit, which, among other activities promotes and commercializes CSIC Intellectual Property to market.

There are several steps that should be complied with for a scientific result to be transferred from the lab to the market, giving rise to a new product, process or service. Based on several cases from CEBAS-CSIC, here I will illustrate factors to consider for taking science from bench to market. From the academic perspective a rational management of the IP portfolio is essential for successful transfer and might be consider as a main limiting factor. IP management should include a clear and monitored internal invention disclosure procedure, a rational evaluation and a professional filling decision committee which will led to the filling of patents on inventions, reasonably strong and with freedom to operate. Having a strong patent is an important factor, but there are other factors that would determine the feasibility of the transfer from bench to market and should be anticipated from the Academia side. These factors include regulatory aspects, market barriers, transaction cost, and other uncertainties that shape the process and the risk of market failure. Technology Transfer experts have here a fundamental role by applying their experience and expertise to analyze and evaluate the risk and possibilities, researching on potential regulatory constrains, searching for the proper partners capable of entering the market, reducing transaction costs and fostering the setting up of a private partner to develop the invention and place it on the market.

Cold pressed plant oils are of increasing popularity among consumers nowadays. They are an attractive alternative to the most popular refined oils and used mainly for salads. Their sensory properties are appreciated together with natural antioxidants and very often a valuable polyunsaturated fatty acids composition. Such oils as pumpkin seeds oil, argan oil, rapeseed and linseed oils are consumed in increasing quantities in recent years.

These oils have often very distinct flavours that originate from the plants oils were pressed from and technological operations (seeds roasting prior to pressing). As fatty acids composition of these oils often favours oxidation processes, flavour compounds being autoxidation products influence these oils sensory properties. Linseed oil is a good example of highly unsaturated oil, which undergoes rapid autoxidation, which results often in consumers rejection of it.

The lecture will be focused on formation of compounds during autoxidation. Profiling of volatile/flavour compounds of different cold pressed oils (fresh and rancid) using comprehensive gas chromatography-mass spectrometry (GCxGC-MS) will be discussed. The use of gas chromatography – olfactometry (GC-O) for the identification of cold pressed oils key odorants and markers for autoxidation will be conferred.

Biodegradable Polymers and Bacteriophages and Their Potential to Guard the Food Safety

We have created a wide family of biodegradable polymers composed of naturally occurring amino acids and other nontoxic building blocks such as fatty diols and dicarboxylic acids. The polymers were synthesized via polycondensation methods of synthesis without using any toxic catalyst or reagents, showed better biocompatibility compared to known biocompatible polymers including poly(lactide/glycolide) copolymers, and are highly promising for numerous biological applications including food processing and safety, especially taking into account that they release α-amino acids upon the biodegradation. The new polymers form micro and nanoparticles that can be loaded by bacteriophages and other biologicals that are applicable to protect perishable foods and beverages.

We have also developed a new facile and cost-effective technology for preparing dry (powdery) bacteriophages using insoluble inorganic absorbents that were add to liquid phages. After filtration or centrifugation of the obtained suspension and subsequent drying of the solid, powdery bacteriophage preparations by 70-75% free of admixtures were obtained. The degree of phage purification can be increased up to at least 99% by repeating desorption-adsorption-filtration process. The obtained powdery phages are ready either to be used directly as food protecting additives or for loading biodegradable micro and nanospheres to be used for the same purposes.

Georgia in spite of small area consists from 15 different soil and up to 30 soil-climatic zones; there are met all types of climate, the only exception is tundra. Being the youngest mountains in Europe, southern slopes of Caucasus are rich by organics. Extraordinary biodiversity of all types of organisms, above 1000 species of endemic plants, collections of all taxonomic groups of microorganisms, accounting above 10.000 strains, animals and insects characteristic for the area of southern Caucasus, existence of the great diversity of genes are making Southern Caucasus area extremely interesting for agricultural, industrial, ecological and pharmaceutical biotechnologies.

Number of plants and microbial based biotechnologies such as: production of stable enzymes from microorganisms extremophiles, development of biofuel production technology, creation of probiotic preparations for human and animals, phage based preparations for biological control in agriculture, microbial production of biosurfactants, production of natural colorants for food industry processing, etc., are currently under development in collaboration with world known companies : Du Pont, Diversa, Pioneer, Sud Chemie, etc., “PhageBioDerm” is biocomposite material based on biodegradable (poly(ester amide) matrix and containing complex of bacteriophages against 5 general suppurative microorganisms (Pseudomonas, E.coli, Staphylococcus, Streptococcus, Proteus) as bactericidal substances, and acting in drug sustained/controlled release fashion. “PhageBioDerm” contains also antibiotic ciprofloxacin, painkiller Benzocain, proteolytis enzyme (chymotrypsin or trypsin). On the base of local vegetative raw materials “NEOPHARM” produces solid, liquid and soft medicinal preparations.

Odours and odour impressions both in a positive and in a negative way have drawn the attention of mankind since ancient times. Not only the positive changes of food smell by the cooking process but also the smell of certain plants and oils have played an important role for centuries.

The odour active substances belong to the volatile fraction of a food product. This is normally the smallest part of the product (usually in the milligram per kilogram range, except in spices where the volatile fraction can be up to five percent of the whole product), nevertheless this is the driving part which is responsible for the acceptance or rejection by consumers.

Off-flavours and taints are defined as unpleasant odours or tastes, the first resulting from the natural deterioration of a food, the second from its contamination by some other chemical.

Although the mass range of odour active substances is limited to 300 Dalton there are literally hundred or thousand compounds having different structure, polarities and hetero atoms, so the chances of spectroscopic and chromatographic interferences is quite likely. Another restriction of analytical approaches is the fact that the method must reach the limits of detection which should be in the range of the sensory threshold. This range spans several orders of magnitude from nanogram per kilogram (or even below) to the upper milligram per kilogram range. In this presentation strategies for the determination of odours and off-odours will be discussed. Presenting several examples for pleasant and unpleasant smell in food products should demonstrate the problems and the approaches to get reliable results. Beside the sample preparation steps the focus on this presentation will be given on the separation and the detection of the target substances.

Activation of 3-O-(3,4,5-Trimethoxybenzoyl)-(-)-Epicatechin (TMECG) in Melanoma to a Potent Irreversible Inhibitor of Dihydrofolate Reductase

Human melanoma is a significant clinical problem because it is resistant to treatment by most chemotherapeutic agents, including antifolates. It is therefore a desirable goal to develop a second generation of low-toxicity antifolate drugs to overcome acquired resistance to the prevention and treatment of this skin pathology. In our efforts to improve the stability and bioavailability of green tea polyphenols for cancer therapy, we synthesized a trimethoxy derivative of epicatechin-3-gallate, which showed high antiproliferative and pro-apoptotic activity against melanoma. This derivative, 3-O-(3,4,5-trimethoxybenzoyl)-(-)-epicatechin (TMECG), is a prodrug that is selectively activated by the specific melanocyte enzyme tyrosinase. Upon activation, TMECG generates a stable quinone methide product that strongly inhibits dihydrofolate reductase in an irreversible manner. The treatment of melanoma cells with TMECG also affected cellular folate transport and the gene expression of DHFR, which supported the antifolate nature of this compound. In addition, its pharmacological efficacy has been confirmed in a mouse melanoma model, in which tumor growth and metastasis were inhibited, significantly enhancing the mean survival of the treated groups. TMECG, therefore, shows a potential for clinical use in melanoma therapy.

Viruses are particularly suitable for applications in nanobiotechnology because their structural components are in the nanoscale range and they have the intrinsic characteristic of self-assembly. In my talk I will describe specific features of several hyperthermophilic viruses that could be of special interest for applications in nanobiotechnology, due to their high stability in diverse extreme conditions and solutes. The viruses have been isolated from hot acidic springs, (> 80°C and pH < 3) in different regions of active volcanism, and infect strains of the hyperthermophilic genera Sulfolobus, Acidianus, Pyrobaculum, and Aeropyrum from the third domain of life, the Archaea.

Natural Remediation Technologies Potential for Production of Safe Food

The constant increase of environmental pollution is one of the most important and unsolved global problems. The sources of chemical pollution are natural and anthropogenic. As a result of urbanization, the unpredictable growth of industry and transport, the annual increase of chemical production, military activities, huge amounts of hazardous substances or toxic intermediate products of their incomplete transformations are accumulated in the biosphere. Bio and phytoremediation technologies are the most reliable, ecologically friendly and cost-effective technologies for rehabilitation of polluted ecosystems.

Plants are equipped with remarkable metabolic and absorption capabilities, as well as transport systems that allow them to uptake and transform environmental organic pollutants into nontoxic compounds or carry out their complete mineralization. Phytoremediation exploits plant physiological processes to decontaminate agricultural soils, industrial sites, brownfields, sediments and water or to improve food chain safety by phytostabilisation of toxic elements.

Knowledge on the detoxification mechanisms in plants, their interactions with microorganisms, combined with genetic analysis and plant improvement, is essential to understand the fate of contaminants in plants and food, nonfood and technical crops. The integration of physicochemical and biological understanding allows the optimization of these processes, making phytotechnologies more effective, decreasing the level and transfer of contaminants along the food chain and augmenting the content of essential minerals in food crops.

Fundamental aspects of pollutants uptake, translocation and transformation in plants, enzyme systems involved in these processes are described.

The production of healthy, nutritious foods is a central theme of the Grand Challenges for Global Health. Research on the relationships between food and diet, the gut microbiota and long-term health is at an all-time high, and the potential for improved nutrition to provide for a long and healthy life for the majority of the world’s population is at the core of many scientific research funding bodies across the developed and developing world.

In many areas, from probiotic bacteria to GM-crops, there are potential products in development that could significantly improve long-term health and wellbeing in the developed world or improved survival and quality of life in the developing world. However, in spite of substantial public attention and research funding, and the huge potential impact that such biotechnological innovation in foods could have on global health, there has been little direct impact to date on the food industry.

I will provide a personal perspective on the reasons for this disconnect between research and application and outline a few of the key obstacles that need to be addressed, before biotechnology can truly make an impact upon our diet.

Mushrooms have been valued for centuries as food or source of pharmacologically-active compounds. A number of bioactive compounds have been isolated from different mushroom species. Many cultures worldwide discovered the medical value of extracts from certain mushrooms and included them as essential components in traditional medicines. On the other hand in last several decades, their therapeutical potential is thoroughly investigated in many laboratories worldwide. Over 300 species are known to possess some therapeutical properties. The most intensively investigated are species of Lentinula, Hericium, Grifola, Flammulina, Pleurotus, Ganoderma, Trametes, and Cordyceps. They exhibit immunomodulatory and anti-cancer activity, anti-diabetic and anti-holesterolemic activity, antimicrobial, antiviral, antiallergic, hepatoprotective and central nervous system-modulating activities. The responsible compounds for bioactivity belong to various chemical groups, from polysaccharides to di- and triterpenes and alkaloids or even complex proteins. In some countries are medicinal mushrooms recognized as valuable functional food, but predominantly they are used as a source for dietary supplements or medicines.